动物造模,缺血再灌注肾损伤动物模型 z-bio 2021-03-26 935

　　(1) Breeding method 30 mg/kg body weight of pentobarbital sodium was injected intraperitoneally into male Wistar rats weighing 200 to 220 g, and fixed on the operating table in the supine position. The animal’s abdominal surgery area was disinfected and depilated regularly, an incision of about 2 cm was made in the lower abdomen, the left renal artery was isolated behind the abdomen, and the left renal artery was clamped with a complete arterial clamp for 60 minutes. After that, the perfusion was restored, and the right kidney was taken out at the same time. The perfusion time (1, 3, 6, 24 hours) was divided into 4 groups. The treatment group was intravenously injected 5 minutes before reperfusion, the sham operation group (without obstruction of renal blood flow) was used as a control, and blood and kidneys were collected at the corresponding time points. An automated biochemical analyzer can measure the levels of serum urea nitrogen (BUN) and creatinine (Cr). The conventional paraffin sections of the kidney tissue were stained with HE, and the pathological changes of the kidney tissue were observed with an optical microscope.

　　(2) Model characteristics After ischemia-reperfusion (IR), P-selectin is significantly expressed in rat kidney tissue, MDA content increases, SOD activity decreases, and obvious cell apoptosis and histopathological changes appear. 24 hours after reperfusion, the serum BUN and Cr of the experimental group were significantly higher than those of the sham operation group. After 1 hour of reperfusion, under an optical microscope, the renal cortex of the experimental group turned white, renal medulla turned black, and renal tubular epithelial cells swelled. Degeneration and necrosis, interstitial congestion and edema, and inflammatory cell infiltration. After the renal blood flow is blocked and the perfusion is restored, the renal tissue will have decreased renal function and obvious pathological changes. Renal tubule epithelial cells were obviously swollen, with varying degrees of degeneration and necrosis, and renal interstitial congestion and edema were related to inflammatory cell infiltration. After ischemia and reperfusion, the kidney will produce a large amount of oxygen free radicals, consume endogenous antioxidants, significantly reduce its ability to scavenge oxygen free radicals, and damage the kidneys. In addition, the production of a large amount of oxygen free radicals can lead to cell DNA denaturation, protein oxidation, lipid peroxidation and even cell death, which is also a factor leading to cell apoptosis during ischemia-reperfusion.

　　(3) Comparative medicine Ischemia-reperfusion injury is very common clinically, but its pathological damage mechanism is still unclear. In recent years, adhesion molecules and their mediated adhesion of leukocytes and endothelial cells are considered to be important factors of ischemia-reperfusion injury. Inhibition of neutrophil infiltration and aggregation by adhesion molecules has been shown to prevent or reduce organ damage caused by ischemia-reperfusion. The animal model of ischemia-reperfusion nephropathy can also be used for adult Japanese white rabbits weighing 2-3 kg. The rabbit was anesthetized by injecting 1% sodium pentobarbital sodium 2.5ml/kg body weight through ear vein, the abdomen was opened aseptically, the left kidney was taken out, and the right renal artery was clamped for 1 hour. Remove to restore blood reperfusion. The animals were comatose 48 hours after the operation, blood was collected from the inferior vena cava, and the serum Cr content was measured by picric acid colorimetry. The animal model of ischemia-reperfusion nephropathy can also be used in isolated pig kidneys infused with autologous whole blood. These kidneys donated 50-80 kg of kidney minipigs. All donor pigs were shocked and braked. After opening the abdomen of the control group, a small part of the kidney was taken and placed in an ice box for testing. The renal cortex of the ischemic group was collected 50 minutes after renal ischemia, and the detection was the same as that of the control group. The experimental group used an isolated pig kidney ischemia-reperfusion injury model. Plasma was collected at 0, 0.5, 1.5, and 2.5 hours after reperfusion, and LDH enzyme activity was measured.